Production of hexosamines and uses thereof
AssigneePharmacia Corporation
InventorMark G. Obukowicz
Life of this patent
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Abstract
A method of producing a hexosamine comprises providing a cell having genes encoding each enzyme required for a biosynthetic pathway capable of synthesizing the hexosamine where at least one gene in the pathway is a heterologous gene. Compositions and methods of producing transgenic cells, expression vectors, transgenic plants, and nutritional material that contain hexosamines are also provided, as are methods for preventing, treating, and inhibiting arthritis and articular-joint damage or disease in subjects in need of such prevention, treatment and/or inhibition.
Claims60
A method for producing a hexosamine comprising:
- providing a cell comprising polynucleotide sequences which encode for enzymes required for a biosynthetic pathway capable of synthesizing the hexosamine, where at least one of the polynucleotide sequences comprises a recombinant polynucleotide.
A method of producing a hexosamine comprising:
- transforming a cell with at least one heterologous polynucleotide coding for a polypeptide in a biosynthetic pathway that is capable of producing a hexosamine;
- and
- culturing the transformed cell under conditions that permit the cell to translate the polynucleotide into a polypeptide comprising an enzyme which is part of the biosynthetic pathway.
A method of producing a hexosamine by a living cell comprising:
- constructing at least one heterologous vector encoding for at least one gene encoding a polypeptide that functions as an enzyme in a hexosamine biosynthetic pathway, wherein the gene is a polynucleotide operably linked to a regulatory promoter sequence;
- transforming at least one cell with the heterologous vector;
- culturing the cell under conditions which allow for expression of the polynucleotide into the polypeptide;
- and
- permitting the cell to produce the hexosamine.
A method of producing hexosamines in transgenic plants comprising:
- constructing at least one heterologous vector encoding for at least one gene in a hexosamine biosynthetic pathway and one gene for a plant selectable marker;
- linking operably the hexosamine pathway gene and the plant selectable marker gene to at least one regulatory promoter that controls expression of said genes, wherein the expression of the genes is capable of producing a polypeptide for enzymatic hexosamine production and a polypeptide for the selectable marker;
- transforming a plant cell with either one or both of the genes operably linked to the at least one regulatory promoter;
- culturing the transformed plant cell under conditions that allow for the plant cell to regenerate into a plant;
- and
- growing the transformed plant under conditions that allow for the plant to produce the hexosamine.
A transgenic plant that is capable of producing a hexosamine, the plant comprising at least one heterologous vector encoding for at least one gene in a hexosamine biosynthetic pathway, wherein the gene is linked operably to at least one regulatory promoter that controls expression of the gene.
A recombinant expression system capable of expressing in host cells either one or both of:
- a gene encoding for a glutamine:fructose-6-phosphate amidotransferase enzyme comprising a polynucleotide sequence that is substantially similar to SEQ ID NO:
- 1;
- and
- a gene encoding for a phosphatase enzyme comprising a polynucleotide sequence that is substantially similar to SEQ ID NO:
- 13,
- both genes being operably associated with a regulatory promoter sequence that controls expression of the genes, wherein the expression system is capable of producing glucosamine in the host cells.
A method of treating a disease in a mammal comprising administering to the mammal by genetic therapy a recombinant expression vector capable of expressing in the mammal at least one least one gene in a hexosamine biosynthetic pathway, wherein the gene is linked operably to at least one regulatory promoter that controls expression of said the gene.
An edible material comprising a hexosamine produced from a recombinant source.
An edible material comprising plant material containing a hexosamine, wherein the plant is a transgenic plant that has been engineered to produce hexosamines.
A method of producing glucosamine from a transgenic plant, the method comprising:
- transforming a cell of the plant with a nucleic acid molecule having a heterologous nucleotide sequence substantially similar to SEQ ID NO:
- 1 encoding a glutamine:fructose-6-phosphate amidotransferase enzyme and a nucleic acid molecule having a heterologous nucleotide sequence substantially similar to SEQ ID NO:
- 13 encoding a phosphatase enzyme, both nucleic acids being operably linked to a regulatory promoter element;
- culturing the transformed plant cell under conditions which allow the plant cell to regenerate into a plant;
- growing the plant under conditions which allow the heterologous nucleic acid molecules to express the glutamine:fructose-6-phosphate amidotransferase and glucosamine-6-phosphatase enzymes;
- allowing the heterologous enzyme glutamine:fructose-6-phosphate amidotransferase to convert fructose-6-phosphate into glucosamine-6-phosphate and the heterologous enzyme glucosamine-6-phosphatase to convert the glucosamine-6-phosphate into glucosamine;
- extracting the glucosamine from the plant;
- and
- purifying the extracted glucosamine into a pharmaceutically accepted form.
A transgenic host cell that is capable of producing a hexosamine, the cell comprising a cell having genes encoding each enzyme required for a biosynthetic pathway capable of synthesizing the hexosamine where at least one gene in the pathway is a heterologous gene.
A method for the prevention, treatment and/or inhibition of arthritis or articular joint damage or disease in a subject in need of such prevention, treatment, and/or inhibition, the method comprising administering to the subject a hexosamine-containing material which comprises polynucleotide sequences which encode for enzymes required for a biosynthetic pathway capable of synthesizing the hexosamine, where at least one of the polynucleotide sequences comprises a recombinant polynucleotide.
Dependencies are inferred from the claim text.